System-Dependent Ecotoxicological Effects of Anatase and Rutile Titanium Dioxide Nanoparticles Across Prokaryotic and Eukaryotic Test Models

The use of nanoscale materials has expanded rapidly in recent decades, with titanium dioxide (TiO2) nanoparticles among the most widely produced. Their increasing environmental release raises concerns about ecosystem-level effects. A key mechanism of toxicity is the generation of reactive oxygen species (ROS), although these effects strongly depend on particle properties, including crystalline form, size, morphology, surface characteristics, and exposure conditions. Here, we assessed and compared the ecological impacts of anatase and rutile TiO2 nanoparticles across prokaryotic and eukaryotic systems, including aquatic microbial communities, microbial cultures, plant bioassays (Sinapis alba, Triticum aestivum), the SOS Chromotest, and zebrafish (Danio rerio) assays. Nano-TiO2 exposure markedly restructured freshwater microbial communities by suppressing sensitive taxa (e.g., Actinomycetota, Flavobacterium, and Limnohabitans) while enriching more tolerant genera such as Pseudomonas, Sediminibacterium, Haliscomenobacter, and Hydrogenophaga. These shifts likely reflect differences in cell-envelope structure, biofilm formation, and antioxidant capacity. The two investigated TiO2 nanoparticle types showed distinct biological effects: rutile was associated with more pronounced microbial community shifts and bacterial cell damage, whereas anatase caused stronger responses in plant assays, particularly by impairing hypocotyl growth and plant water balance. Besides the limited acute genotoxicity revealed by the SOS Chromotest, TiO2 nanoparticles did not significantly affect survival or final larval body length in the zebrafish embryo assay under the tested conditions; however, the hatching delay observed at the highest concentration indicated a sublethal developmental effect. Overall, our results show that TiO2 nanoparticle toxicity cannot be generalized across biological systems and suggest that biological responses depend on the combined influence of particle characteristics and organism-specific structural and physiological traits.